This Alcohol Research Center Grant (ARC) is focused on the unifying hypothesis that common cellular and molecular events caused by ethanol lead to alterations in cellular signaling that trigger tissue specific pathologies. Interdisciplinary research themes include studies on cellular signaling, calcium levels, steroids, gene regulation, transcription factor activation, oxidative stress behavioral responses to ethanol and the use of gene delivery to investigate and/or modify the effects of ethanol. The ability to study single cell types in vitro complements and strengthens studies of molecular signaling events in specific brain regions. The pathologies associated with alcoholism represent a central theme that is divided into two foci, tissue pathology and the pathological processes that regulate alcohol seeking behaviors. This ARC connects 15 funded research projects focused on molecular and cellular processes that underlie the pathogenesis of alcoholism and alcohol toxicity. Oxidative tissue damage is a focus of several components and cores that link eight currently funded research projects together within this Center to test the hypothesis that oxidative stress is a major mechanism of ethanol toxicity. Components within the ARC will study free radical production, cytosolic calcium signaling, mitochondrial calcium and membrane potential, activation of transcription factors NFkappaB and APl, cytokine formation and the activation of kinase signaling cascades. The interaction of CYP2EI, reperfusion-oxygen tension and other determinants of oxidative stress will be related to apoptosis and cell death. Factors that determine gene expression and transcription factor activation will connect liver, brain, fetus and pancreas. Three components of this ARC will focus on brain and bridge together seven active research projects studying the cellular and molecular mechanisms that promote addictive behavior. The molecular mechanisms that control brain reward systems and ethanol withdrawal sensitization will be studied to elucidate the pathogenesis of alcohol addiction in brain. The signaling pathways involved in these processes will be determined by following transcription factor formation, induction of immediate early genes, regulation of ethanol sensitive receptor genes and the ability of steroids and delivered genes to modulate ethanol self administration, preference and dependence. Gene delivery will be used to modify transcription factor activation and GABAa receptor sensitivity as well as to deliver antioxidants and enzymes that enhance neurotransmitter synthesis. Models of chronic ethanol administration, eg. Tsukamoto-French, repeated ethanol withdrawal, chronic self administration, will be shared among components providing greater insight into the aspects of ethanol consumption that contribute to toxicity. Studies of both males and females are included in several components to better understand the role of gender and steroid hormones m ethanol pathology, particularly since steroids appear to modulate ethanol dependence and the free radical capacity of cells. These studies build on existing knowledge in different tissues and cellular systems that allow molecular and cellular hypothesis on ethanol toxicity to be specifically tested. Studies in multiple cellular and tissue systems strengthen the ability of investigators to explore new avenues to uncover the molecular mechanisms of alcohol pathogenesis.